Maleimide Activated Agarose


Agarose Guide
PureCube Maleimide Activated Agarose is available as off-the-shelf item and as a bulk product. Our careful in-house production provides low lot-to-lot variation, making activated agaroses also suitable for commercial production. PureCube Maleimide Activated Agarose carries a maleimide functionality. Proteins bind covalently and irreversibly to free thiol groups.

PureCube Maleimide Activated Agarose provides:


Activated Maleimide agarose resins from Cube Biotech were successfully used in the following publications and patents:

Purified SubstanceYearAuthor
cap-specific nucleoside 2′-O-methyltransferase. 2016 Roos T., Yazdan Oanah B., Conzelmann M., Thess A., Buob D., Kunze M., Wagner V.2016-August-121
Recombinant mutant PPases 2018 Kunze M., Hadler F.N., Yazdan Panah B.,Ross T. 2
P2-GM1os 2019 Mahon C.S., Wildsmith G.C., Haksar D., de Poel E., Beekman J.M., Pieters R.J., Webb M.E., Turnbill W.B.3

Features

Usage Coupling proteins to via their activated thiol groups
Specifity Affinity to thiol groups
Flow Rate up to 6 mL/min
Filling quantity Delivered as a 50 % suspension
Bead size 40 µm
Bead Ligand Affinity agarose with activated Maleimide groups
Required equipment
 
  • Coupling Buffer I (PBS) pH 7.2, 250 mL
  • Coupling Buffer II pH 6.5, 250 mL
  • Storage Buffer I, pH 6.5, 250 mL
  • Storage Buffer II, pH 7.5, 250 mL
  • Centrifuge for 15 mL tubes
  • Centrifuge tubes (15 mL)
  • End-over-end mixer or thermomixer
  • Spectrophotometer

Applications

All protocols and buffer compositions are also avaible as PDF-Files on the Protocols & Datasheets page.
   
A.Maleimide coupling protocol
 
 
  1. Transfer 1 mL PureCube Maleimide-Activated Agarose suspension (corresponding to 1 mL bed volume) into a 2 ml microcentrifuge tube.Tip: The coupling reaction can be linearly scaled up and down, by increasing or decreasing the amounts of buffers and solutions described in this protocol.
  2. Spin the tube at 500 x g to pellet the agarose. Remove the supernatant.
  3. Wash the Activated Agarose three times with 1 ml coupling buffer I or II. Tip: Choose coupling buffer I or II depending on the pH stability of your protein. Use the same coupling buffer throughout the procedure.
  4. Resuspend 1-3 mg protein in 500 µl coupling buffer I or II.
  5. Add the protein to the activated agarose and mix by vortexing. Tip: When coupling a particular protein for the first time, try 3-5 different protein concentrations to make sure you are offering enough protein in the reaction but not wasting any protein.
  6. Depending on the temperature stability of the protein, incubate at 20°C for 2 hours or at 4° C overnight on an endover-end shaker or thermoshaker.
  7. Spin the tube at 500 x g to pellet the agarose. Remove the supernatant and analyze the supernatant in a spectrophotometer. Record absorption at 280 nm to monitor coupling efficiency.
  8. Wash the agarose three times with 1 ml coupling buffer and once with 1 ml double distilled water.
  9. Tip: Monitoring absorbance at 280 nm tells you about the coupling efficiency of the protein (compare A280 of the original protein solution to the supernatant in step 6 to determine % coupling). It also helps you identify the optimal amount of protein required for efficient coupling.
  10. Resuspend the coupled Agarose in 1 mL Storage buffer I or II, yielding a 50% suspension. Store at 4°C.

References

1. Roos T., Yazdan Oanah B., Conzelmann M., Thess A., Buob D., Kunze M., Wagner V. inventor; Method for adding cap structures to rna using immobilized enzymes. International patent: WO2016193226A1. 2016-December-08
2. Kunze M., Hadler F.N., Yazdan Panah B.,Ross T. inventor; CureVae AG, Tübingen, Immobilized Inorganic Pyrophosphatase (Ppase), Us-Patent: US 2019 / 0177714 A1; September-21-2018
3. Mahon S. et al. (2019). A ‘catch-and-release’ receptor for the cholera toxin. Faraday Discussions. 10.1039/C9FD00017H.
Agarose Guide
PureCube Maleimide Activated Agarose is available as off-the-shelf item and as a bulk product. Our careful in-house production provides low lot-to-lot variation, making activated agaroses also suitable for commercial production. PureCube Maleimide Activated Agarose carries a maleimide functionality. Proteins bind covalently and irreversibly to free thiol groups.

PureCube Maleimide Activated Agarose provides:


Activated Maleimide agarose resins from Cube Biotech were successfully used in the following publications and patents:

Purified SubstanceYearAuthor
cap-specific nucleoside 2′-O-methyltransferase. 2016 Roos T., Yazdan Oanah B., Conzelmann M., Thess A., Buob D., Kunze M., Wagner V.2016-August-121
Recombinant mutant PPases 2018 Kunze M., Hadler F.N., Yazdan Panah B.,Ross T. 2
P2-GM1os 2019 Mahon C.S., Wildsmith G.C., Haksar D., de Poel E., Beekman J.M., Pieters R.J., Webb M.E., Turnbill W.B.3

Features

Usage Coupling proteins to via their activated thiol groups
Specifity Affinity to thiol groups
Flow Rate up to 6 mL/min
Filling quantity Delivered as a 50 % suspension
Bead size 40 µm
Bead Ligand Affinity agarose with activated Maleimide groups
Required equipment
 
  • Coupling Buffer I (PBS) pH 7.2, 250 mL
  • Coupling Buffer II pH 6.5, 250 mL
  • Storage Buffer I, pH 6.5, 250 mL
  • Storage Buffer II, pH 7.5, 250 mL
  • Centrifuge for 15 mL tubes
  • Centrifuge tubes (15 mL)
  • End-over-end mixer or thermomixer
  • Spectrophotometer

Applications

All protocols and buffer compositions are also avaible as PDF-Files on the Protocols & Datasheets page.
   
A.Maleimide coupling protocol
 
 
  1. Transfer 1 mL PureCube Maleimide-Activated Agarose suspension (corresponding to 1 mL bed volume) into a 2 ml microcentrifuge tube.Tip: The coupling reaction can be linearly scaled up and down, by increasing or decreasing the amounts of buffers and solutions described in this protocol.
  2. Spin the tube at 500 x g to pellet the agarose. Remove the supernatant.
  3. Wash the Activated Agarose three times with 1 ml coupling buffer I or II. Tip: Choose coupling buffer I or II depending on the pH stability of your protein. Use the same coupling buffer throughout the procedure.
  4. Resuspend 1-3 mg protein in 500 µl coupling buffer I or II.
  5. Add the protein to the activated agarose and mix by vortexing. Tip: When coupling a particular protein for the first time, try 3-5 different protein concentrations to make sure you are offering enough protein in the reaction but not wasting any protein.
  6. Depending on the temperature stability of the protein, incubate at 20°C for 2 hours or at 4° C overnight on an endover-end shaker or thermoshaker.
  7. Spin the tube at 500 x g to pellet the agarose. Remove the supernatant and analyze the supernatant in a spectrophotometer. Record absorption at 280 nm to monitor coupling efficiency.
  8. Wash the agarose three times with 1 ml coupling buffer and once with 1 ml double distilled water.
  9. Tip: Monitoring absorbance at 280 nm tells you about the coupling efficiency of the protein (compare A280 of the original protein solution to the supernatant in step 6 to determine % coupling). It also helps you identify the optimal amount of protein required for efficient coupling.
  10. Resuspend the coupled Agarose in 1 mL Storage buffer I or II, yielding a 50% suspension. Store at 4°C.

References

1. Roos T., Yazdan Oanah B., Conzelmann M., Thess A., Buob D., Kunze M., Wagner V. inventor; Method for adding cap structures to rna using immobilized enzymes. International patent: WO2016193226A1. 2016-December-08
2. Kunze M., Hadler F.N., Yazdan Panah B.,Ross T. inventor; CureVae AG, Tübingen, Immobilized Inorganic Pyrophosphatase (Ppase), Us-Patent: US 2019 / 0177714 A1; September-21-2018
3. Mahon S. et al. (2019). A ‘catch-and-release’ receptor for the cholera toxin. Faraday Discussions. 10.1039/C9FD00017H.
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Maleimide Activated Agarose
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PureCube Maleimide Activated Agarose PureCube Maleimide Activated Agarose
2 ml 50% Maleimide-Activated Agarose, for direct, irreversible coupling of biomolecules via thiol groups For more information read here:...
Article number: 51103
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